Anti-Deinagkistrodon acutus venom properties of ethanolic root extract from Cynanchum paniculatum (Bunge) kitag and its GC-MS analysis

Abstract

Ethnopharmacological relevance: Cynanchum paniculatum (Bunge) Kitag known as a popular Chinese herbal medicine has been used for a long time to treat a wide variety of diseases including snakebites. However, there is scarce information on the antiophidian potential of this plant.

Aim of the study: The purpose of this work was to evaluate the inhibition effects of the ethanol extract of C. paniculatum on the enzymatic and biological activities induced by Deinagkistrodon acutus venom (DAV). The phytochemical components in the extract were also determined for understanding the mechanism of antivenom activities.

Materials and methods: Fresh root of C. paniculatum was shed-dried and smashed into powder. The powder was then extracted in 75% ethanol by refluxing method. Inhibition of proteolytic, phospholipase A₂, fibrinogenolytic and hyaluronidase activities of DAV by the extract were determined in vitro. Neutralization of lethal, hemorrhagic, myotoxic and edematogenic activities induced by the venom were also performed in vivo. Phytochemical constituents of the extract were analyzed by gas chromatography-mass spectrometry (GC-MS).

Results: The ethanolic root extract of C. paniculatum (CPER) was able to completely (100%) inhibit protease and hyaluronidase activities induced by DAV when preincubated at a ratio of 1:100 and 1:50 (venom/extract, W/W), respectively. PLA₂ and fibrinogenolytic enzyme activities were actually neutralized at the concentration tested. In-vivo studies the inhibition of venom hemorrhagic and myotoxic action reached 100% when the venom was previously incubated with the extract (1:100) before injection. The edematogenic effect was also inhibited in a dose-response manner. CPER completely inhibited DAV-induced lethality in mice when the venom was preincubated with the extract at a ratio of 1:100. GC-MS analysis indicated that a total of 58 compounds were discovered in CPER. Among them, 5 bioactive constituents including 9,12-octadecadienoic acid (Z,Z)-, n-hexadecanoic acid, cis-vaccenic acid, γ-sitosterol and stigmasterol exhibit antivenom activity according to previous reports.

Conclusions: The results obtained in this work validate for the first time CPER as a traditional antiophidic herb, especially inhibitory effect on local damage induced by DAV, suggesting it could be used as a potential source of bioactive components against snakebites.

Keywords: Cynanchum paniculatum (Bunge) Kitag; Deinagkistrodon acutus; Ethanolic root extract; GC-MS; Venom.